Cannon R E, White J A, Scandalios J G
Proc Natl Acad Sci U S A. 1987 Jan;84(1):179-83. doi: 10.1073/pnas.84.1.179.
A cDNA clone encoding maize cytosolic superoxide dismutase 2 (SOD2) was isolated from a cDNA library constructed in pUC9 plasmids from size-selected poly(A)+ RNA. The library was screened with mixed synthetic oligode-oxynucleotide probes. The sequence of the probes was derived from the amino acid sequence from a region of the protein near the NH2 terminus. One positive clone contained an insertion of a 612-base-pair fragment. The amino acid sequence, deduced from the nucleotide sequence of the cDNA, revealed that the clone contained the coding region for all but the first of the 151 amino acids of the SOD2 protein. Additional 5' and 3' flanking sequences, absent on the pUC9 Sod2 clone, were obtained from a lambda gt11 clone isolated from a maize leaf library probed with the pUC9 Sod2 insert. Hybrid-selection translation assays also demonstrate that the cDNA clone contains Sod2 sequences.
从以pUC9质粒构建的、经大小选择的聚腺苷酸加尾（poly(A)+）RNA的cDNA文库中分离出一个编码玉米胞质超氧化物歧化酶2（SOD2）的cDNA克隆。用混合合成寡脱氧核苷酸探针筛选该文库。探针序列源自蛋白质靠近氨基末端区域的氨基酸序列。一个阳性克隆包含一个612个碱基对片段的插入。从cDNA的核苷酸序列推导的氨基酸序列表明，该克隆包含SOD2蛋白151个氨基酸中除第一个氨基酸外所有氨基酸的编码区。从用pUC9 Sod2插入片段探测的玉米叶文库中分离出的λgt11克隆获得了pUC9 Sod2克隆上不存在的额外5'和3'侧翼序列。杂交选择翻译分析也证明该cDNA克隆包含Sod2序列。
